PROVIDED. Self antigens are recognized on cancers by the immune system of patients with cancer. Immune responses to self antigens can lead to rejection of cancer in laboratory models. Immunization against these antigens presents problems, however, because these are weak antigens. This proposal addresses limitations of the present molecularly defined cancer vaccines targeting self-antigens: 1) insufficient potency, 2) specificity for only individual epitopes or determinants, and 3) a need to modulate the immune system to favor response to vaccination. Using prototypical differentiation antigens of the tyrosinase family for models, creation of multiple heteroclitic epitopes and manipulation of protein stability and trafficking will be studied to increase potency of active immunization against cancer self-antigens. In the first Aim, DNA vaccines are designed to introduce multiple heteroclitic epitopes and to enhance antigen processing and presentation, followed by testing in laboratory treatment models for induction of antibody and T-cell responses and for antitumor activity. The second Aim investigates strategies to increase immune responses to vaccination through modulation of homeostatic immune cell recovery. Another strategy is explored to immunological target both cancer self-antigens and tumor stroma through active immunization or combination of active and passive immunization. The third Aim investigates immune modulation in combination with DNA vaccination in a clinical study. Finally, the last Aim explores DNA vaccination against cancer self-antigens in combination with vaccination against tumor stroma in a clinical study.